FI80885C - Preparation of new guanine derivatives - Google Patents

Description

1 80885

Manufacture of new guanine derivatives

The invention relates to a process for the preparation of therapeutically active guanine derivatives according to the preamble of claim 1.

Finally, the effects of viruses on bodily functions can lead to changes that occur at the cellular level and below. Pathogenic changes at the cellular level are different depending on the different combinations of viruses and host cells. When a few viruses cause complete damage to certain cells (cell death), a few others can create a neoplastic state in the cells.

Important common viral infections include Herpes dermatitis (this includes Herpes labialis), Herpes keratitis, Herpes genitalis, Herpes zoster, Herpes encephalitis, infectious mononucleosis, and cytomegalovirus infections, all of which are caused by viruses that belong to the group of herpes viruses. Other important viral diseases are influenza A and B, which are caused by influenza A and B viruses, respectively. Other important common viral diseases include viral hepatitis and especially hepatitis B virus infections, which are widespread. There is a need for effective and selective virus-like agents in the treatment of these diseases, as well as in the treatment of other viral diseases.

Several different viruses, both of the DNA and RNA types, have been shown to cause tumors in animals. The action of carcinogenic chemicals can cause the activation of latent tumor viruses in animals. It is possible that tumor viruses are present in human tumors. The most likely cases currently known in humans are leukemia, sarcoma, breast carcinoma, Burkitt's lymphoma, nasal carcinoma, and neck cancer, in which RNA-2 80885 tumor viruses and herpes viruses have been identified. This makes it important to investigate for selective inhibitors of tumorigenic viruses and their effects in cancer resistance efforts.

The following publications have previously described guanine derivatives: Synthetic Communications 2 (6), 345-351 (1972), U.S. Patents 4,199,574 and 4,355,032, European Patent Application 82401571.3 (Publication No. 74,306) and European Patent Application 81850250.2 (Publication No. 55 239), in which the latter discloses antiviral compounds of the formula

O

1 1

CH2-CH-CH-CH 2 OH

I I

R 1 is R 2 wherein R 1 and R 2 may be hydrogen, hydroxy or fluoro.

EP-141 927 discloses antiviral compounds of the following formula

X

l Y> Η2Ν '> ^ Ν' ^ ΚχΝ ^

CH 2 CH 2 CH 2 OH

ch2oh wherein X represents chlorine, alkoxy, phenoxy, phenylalkoxy, -NH2, -OH or -SH. EP-141 927 was filed on August 10, 3,80885 and became public on May 22, 1985, and is thus relevant to novelty but not inventiveness compared to the present invention.

The present invention relates to a method according to the preamble of claim 1.

It has been found that the compounds of the invention have antiviral activity and inhibit certain viral functions, such as tumorigenic functions, and viral replication.

It has also been found that the compounds of the invention have a very high affinity for viral thymidine kinase, which is very important for antiviral activity when these compounds are used in cells rich in thymidine.

The invention thus provides compounds, as well as geometric isomers and physiologically acceptable salts thereof, which compounds are useful in the therapeutic and / or prophylactic treatment of viral diseases and which may be useful in the therapeutic and / or prophylactic treatment of viral cancer.

4 80885

An effective selective antiviral agent with tolerable side effects should have a selective inhibitory effect on the specific effect of the virus being treated. Accordingly, it is an object of the present invention to provide a novel method for controlling viral infections using an antiviral agent which has a selective inhibitory effect on viral functions but has only a negligible inhibitory effect on the function of stem cells.

One very important application of the compounds of the invention is in the treatment of herpes virus infections. Among the herpes viruses, mention may be made of Herpes simplex species 1 and 2, varicella (Herpes zoster), a virus that causes a mononucleosis infection (i.e., Epstein-Barr virus), and a cytomegalovirus. Important diseases caused by herpes viruses include Herpes dermatitis (including Herpes labialis), Herpes genitalis, Herpes keratitis, Herpes enkephalitis, and Herpes zoster.

Another possible use of the compounds of the invention is in the treatment of cancer and tumors, especially those caused by viruses. This effect can be achieved in different ways, i. by preventing the transformation of virus-infected cells into a neoplastic state by preventing the spread of viruses from the altered cells to other normal cells and by preventing the growth of virus-infected cells.

Another field of application of the compounds of the invention is the inhibition of the growth of transformed cells by the use of specific herpes virus enzymes, such as thymidane kinase, in these cells.

Possible areas for the use of the compounds of the invention in the chemotherapy of cancer include the treatment of the following diseases: leukemias, lymphomas, including Burkitt's lymphoma and Hodgkins' disease, sarcomas, breast carcinomas, pharyngeal and nasal carcinomas and neck cancers caused by viruses.

Other possible fields for the use of the compounds of the invention in the chemotherapy of cancer are the treatment of the following diseases: Multiple myeloma and lung cancer (and bronchial ...

cancer), gastric cancer, liver cancer, colon cancer, bladder cancer, lip cancer, bone cancer, kidney cancer, ovarian cancer, prostate cancer, pancreatic cancer, skin cancer (melanoma), colorectal cancer (cancer), cancer of the colon, salivary cancer, lip cancer, lip cancer, , bladder cancer (and urinary tract cancer), nasal cancer ,. laryngeal cancer, tissue cancer, penile cancer, pubic cancer, vaginal cancer, corpus uterine cancer and tongue cancer. -

The invention provides a medicament suitable for use in: A. A method of treating a viral disease in an animal, including a human, comprising administering to a thus infected animal a therapeutically effective amount of a compound of formula I or a physiologically acceptable salt thereof.

B. A method of inhibiting the multiplication of viruses, in particular herpes viruses, in animals, including man, by administering to an animal in need of such treatment a compound of formula I or a physiologically acceptable salt thereof in an amount sufficient to prevent such multiplication. .

C. A method of treating neoplastic diseases caused by a virus in an animal, including a human, by inhibiting the growth of cells in which viral activity is present, thereby administering to the infected animal a therapeutically effective amount of a compound of formula I or a physiologically acceptable salt thereof.

D. A method of inhibiting the growth of virus-transformed cells in an animal, including a human, by administering to an animal in need of such treatment a compound of formula I or a physiologically acceptable salt thereof in an amount sufficient to inhibit this growth.

6 80885 E. A method of treating viral neoplastic diseases in an animal, including a human, by inhibiting the proliferation of tumor viruses by administering to an animal in need of such treatment a compound of formula I or a physiologically acceptable salt thereof in an amount sufficient to inhibit such proliferation.

F. A method of treating neoplastic diseases in an animal, including a human, by administering to such an animal a therapeutically effective amount of a compound of formula I or a physiologically acceptable salt thereof.

As mentioned above, the novel compounds prepared by the process of the invention have the formula

O

61 7

1 HN X8 I

2i 4 L /

HaN / 3 19

A

wherein A is (CHa) m-CH- (CHa) nOH (Aa) or

Ra CHa-C = C-CHaOH (Ab)

I I

Ri Rt, in which m is 1 or 2, n is 1 or 2, where m is 1 when n is 2 and m is 2 when n is 1,

R 1 is (CHa) pOH or COR 1, R a is hydroxy, p is 1-4,

R 1 and R 1, are independently selected from hydrogen and the compound (CH 2 JpOH, provided that at least one of R 1 or R 1 is hydrogen, or R 1 and R 1 together form a carbon-C 80 a-bond to form an alkyne and further provided that when m is 2 and n is 1 and RA is (CH 2 OH, then p is 2-4, or a physiologically acceptable salt, geometric or optical isomer thereof for the preparation of a compound of formula I wherein A is Aa .

The above definitions of m, n and RA mean that the following specific compounds, including their salts and optical isomers, form part of the invention:

O

1 Y>

CH 2 CH 2 CH 2 OH

RA

COOH

ch2ch2oh ch2ch2ch2oh ch2ch2ch2ch2oh o

Lj> h2n ch2-ch-ch2-ch2oh

RA

COOH

CH 2 OH

CH2CH2OH

ch2ch2ch2oh ch2ch2ch2ch2oh 8 80885

The compounds of formula I contain one asymmetric center when R e and (CH 2) n OH are different. Accordingly, they exist in two optical forms, which also form part of the invention.

The following specific compounds, including their salts and geometric isomers, also form part of the invention:

O

tt i T>

H 2 N

CH2-C .................- ‘C-CH2OH

I I

Ri R &

H H

H CH2OH

H CH2CH2OH

H CH2CH2CH2OH

H CH2CH2CH2CH2OH

CHaOH H

CH2CH2OH H

CH2CH2CH2OH H

CH2CH2CH2CH2OH H

When Rt. is not CH2OH, the compound exists as two geometric isomers which also form part of the invention.

The invention comprises the following compounds: 9- [4-hydroxy-2- (hydroxymethyl) butyl] guanine 9- [4-hydroxy-3-carboxybutyl] guanine 9- [5-hydroxy-3- (hydroxymethyl) pentyl] guanine 9 - [6-hydroxy-3- (hydroxymethyl) hexyl] guanine 9- [4-hydroxy-2-carboxybutyl] guanine 9- [4-hydroxy-2- (hydroxyethyl) butyl] guanine 9- [5-hydroxy-2- (hydroxyethyl) pentyl] guanine 9 80885 9- [6-hydroxy-2- (hydroxyethyl) hexyl] guanine 9- [4-hydroxy-3- (hydroxymethyl) -2-butenyl] guanine 9- (4-hydroxy-2-butenyl) guanine cis-9- (4-hydroxy-2-butenyl) guanine 9- (4-hydroxy-2-butynyl) guanine

A preferred subgroup of compounds of the invention is obtained when R 1 is (CH 2> pOH, such as hydroxymethyl or hydroxyethyl).

Other preferred compounds of the invention are 9- [4-hydroxy-3- (hydroxymethyl) -2-butenyl] guanine and cis-9- (4-hydroxy-2-butenyl) guanine.

A compound of formula I in which and R 11 form a bond has antiviral activity, and this compound is also an intermediate of the compounds of formula I in which R 1 and R 2 are hydrogen atoms.

The compounds of the invention may be obtained by any of the following methods A-E, which also form part of the invention.

A. Condensation of the acyl side chain, wherein the functional groups may optionally be protected at the N-9 position of the guanine derivative, followed by removal of the protected groups by one or more chemical reactions.

? 6a I N f * 2a [Y 7 ♦ Xa- (CH2) „, - CH- (CH2) nOR3a ->

4a-N

I I

R5a Y

10 80885

Rea - * jlJTV - *

Ria-N - ^ N ^ NN Rja

I I I

R = a (CHa) m-CH- (CH2) nOR3a 0

And

HaN RA

I I

(CH) m CH (CH 2) n OH

wherein RA, m and n are as defined above, Xa is a leaving group such as chlorine, bromine, iodine or OSOaR7Ä, wherein R7a is alkyl, fluorinated alkyl, arylalkyl or aryl, Y is hydrogen or a quaternary ammonium ion, R3a is hydrogen or a protecting group , such as acyl, alkoxycarbonyl, arylcarbonyl, silyl or SOaR-a, Ra «is RA as defined above, or CN, COORea, CH (OR9e) 2, CH (SR9a) a, NHCONHR9 or (CHaJpORga, where p is as defined above,

Rea is a substituted silyl or a group R 7a as defined above, R 8a is a group R 3a as defined above, or Rea and R 10a together with the group R 3a form a divalent group, i.e. a cyclic derivative such as a lactone, a cyclic ester, an orthoic acid, an acetal, an ether or a silyl-type compound, R 4 «and R 5« are the same as R 3a above, R e «is a leaving group such as Xa, OH or ORxoa as defined above, is alkyl, arylalkyl, SO 2 R 7a, substituted silyl, phosphoryl diester or phosphinothioyl. Protecting groups are well known to those skilled in the art (see, e.g., "Protective Groups in Organic Chemistry", T. W. Greene, Wiley 1981; "Methoden der Organischen Chemie",

Houben-Weyl Vl / lb; and "Comprehensive Organic Chemistry", D.H.R. Barton and W.D. Ollis et al., 1979, Vol. 1, pp. 623-629).

il 80885

Some examples of R 3a are acyl groups such as acetyl or benzoyl, alkoxycarbonyl or aryloxycarbonyl groups, silyl groups such as e.g. tert-butyldimethylsilyl, arylalkyl such as benzyl and triarylmethyl or SO 2 R 7a where Rya is as defined above.

The condensation is preferably carried out in an organic solvent such as dimethylformamide, ethanol, acetone in trilane or dichloromethane at a temperature between 0 ° C and 100 ° C for 1 hour to 3 days in the presence of a base (when Y is H), e.g. potassium carbonate.

After condensation, these compounds are hydrolyzed at 0-100 ° C for 1-24 hours using an acid or base such as acetic acid, hydrochloric acid (1-35%) in water, sodium hydroxide (1-20%) in water, ammonia (1-25%) in water or methanol or hydrogenated with hydrogen gas in an organic solvent such as ethanol or dimethylformamide using a metal catalyst for 1 to 24 hours at a pressure of 100 to 5,000 kPa (1 to 50 atmospheres). When R 2a is CH (SRga) 2, the hydrolysis can be performed using a reagent such as mercuric chloride in a separate step.

12 80885 B. Condensation of an unsaturated side chain in which the functional groups may optionally be protected at the N-9 position of the guanine derivative using a basic catalyst followed by reduction and / or deprotection R 1; 6 a T) 14a

4a | I

R H

R5a R,. 6a

Y / N

-> ilX)> -> 43 I i h 4a R_ CH CH-CH.-R Γ 5a 22 15a —1 ..ÄX.

* | | 16a

CH2-CH2-CH2 OH

R 1a, R 8a and R 8a are as defined above, R 6a is CN, CONH 2 or COOR 8a, and R 15a is COOR 8a or CH 2 OR 9a, wherein R 8a and R 8a are as defined above, and may further form a divalent group, i. a cyclic derivative such as a lactone or other cyclic ester, orthoic acid, acetal, ether or a silyl-type compound, and R 1 ga is CONH 2, COOH or CH 2 OH.

13 80885

The condensation is preferably carried out in an organic solvent such as dimethylformamide, ethanol, acetonitrile or dichloromethane at a temperature between 0 and 100 ° C for 1 hour to 3 days in the presence of a basic catalyst such as potassium carbonate, sodium methoxide, sodium hydride or tetrabutylammonium ammonium.

C. Reduction or selective reduction of derivatized or activated carboxylic acid groups followed by deprotection.

R, fl s - »» ..T ϊ ».

R (CH ~) -CH- (CH9) ..- COR., 2 m 2 n-1 11a R, 6a ___>

R4a-N '^ N ^ \ N' h 3a 4a I II

R_ (CH-) -CH- (CH9) OR ~ 5a 2 m 2 n and in which formulas m, n, R-, R., Rj- and Rc are as defined above, όΟ 4 cl 33 Da R ,, is R, . as defined above, or such a group

1 iu O

as (CH 2) 2 N = CHO-, in an activated carboxylic acid derivative,

R12a is a group R2a (C2) p_] _ COR ^ a, at 3 ° - * R2a '^ a ^ a P is as defined above and R 1a is R 1 or R 2a' as defined above.

The reduction can be carried out using hydrogen gas or hydrogen formed in situ and using a metal as a catalyst, or preferably a hydride reducing agent in an organic solvent.

14 80 8 85 D. An acyl side chain condenser in which the functional groups may optionally be protected at the N-9 position of the guanine derivative, followed by removal of the protecting groups by one or more chemical reactions.

R6b I N R2b R7b

\ I I

Γ | \ + xb - CH2 - c = C - CH2OR3b ->

R4h-N "^. N

I I

R5b Y

R6b - rV '\ - R4b-N- ^ N / \ N / R2b R? B R5b CH2 - C = C - CH2OR3b 0

.OF

-> xo

HoN - "^ \ N N RiL RA

III

ch2 -c = c - ch2oh 80885 wherein Xb is a leaving group such as chlorine, bromine, iodine or OSO2R8b 'wherein R8b is alkyl, fluorinated alkyl, arylalkyl or aryl, Y is hydrogen or a quaternary ammonium ion, R3b is hydrogen or a protecting group such as acyl, alkoxycarbonyl, aryloxycarbonyl, silyl or SO 2 R 9b, R 2b is R e or (CH 2) pOR 9b and R 6b is R 1 or (CH 2) pOR 9b wherein p is 1-4 and wherein R 9b is a group R 3b as defined above or R 9b and R 3b together form a divalent group, i.e. a cyclic derivative such as a carbonate ester, thiocarbonate ester, orthoic acid or acetal compound,

Rgb is hydroxy or a leaving group such as chlorine, bromine, iodine, mercapto, alkylthio, SO 2 Rgb or OR 1 Qb 'wherein R 1b is alkyl, arylalkyl, substituted silyl, phosphoryl diester, phosphinothioyl or SO 2 R gb, R 4b and Rgb are the same as above defined group R3b. R3b may be a hydroxy protecting group well known to those skilled in the art (see, e.g., "Protective Groups in Organic Chemistry", TW Greene, Wiley 1981; "Methoden der Organischen Chemie", Houben-Weyl Vl / lb; and "Comprehensive Organic Chemistry ", DHR Barton and WD Ollis, 1979, Vol. 1, pp. 623-629).

Some examples of R 3b are acyl groups such as acetyl or benzoyl, alkoxycarbonyl or aryloxycarbonyl groups, silyl groups such as e.g. tert-butyldimethylsilyl, arylalkyl such as benzyl and triarylmethyl, or SO 2 R gb is as defined above, wherein R gb is as defined above.

16 80 885

The condensation is preferably carried out in an organic solvent such as dimethylformamide, ethanol, acetonitrile or dichloromethane at a temperature between 0 and 100 ° C for 1 hour to 3 days in the presence of a base (when Y is H), which may be, for example, potassium carbonate.

After condensation, the compounds are hydrolyzed at 0-100 ° C for 1-24 hours using an acid or base such as acetic acid, hydrochloric acid (1-35%) in water, sodium hydroxide (1-20%) in water, ammonia (1-25%) in water or methanol.

E. Condensation of an unsaturated side chain, wherein the functional groups in the side chain may optionally be protected at the N-9 position of the guanine derivative using a metal catalyst followed by deprotection by one or more chemical reactions.

? 6b / N R p K \ (\, 2b, 7b

Li /> + CH = c - C - CH, - * R4b -

R5b H

I 6b _>

Rih · N '^ N r 4b I I? 2b R7b

R5b CH2 - c = c - CH2OH

o / A.

1 u S ·

ch2 - c = C - CH2OH

R ^ t Rb »R2b '^ 4b' R5b 'R6b - * a R7b ° n as defined above.

17 80885

The condensation is preferably carried out in an organic solvent such as dimethylformamide, ethanol, acetonitrile or dichloromethane at a temperature between 0 and 100 ° C for 1 hour to 3 days over a metal catalyst such as tetrakis (triphenylphosphine) palladium ( In the presence of 0).

The disclosed methods A-C can be used to obtain mixtures of optical isomers or, where appropriate, to obtain a single optical isomer. In addition, a single optical isomer can be obtained from racemic mixtures by methods known per se.

The methods D and E described can be used to obtain mixtures of geometric isomers or, where appropriate, to obtain only one geometric isomer. Only one geometric isomer can also be obtained from mixtures of isomers by methods known per se.

The starting materials in the above-mentioned methods A-E are either known compounds or can be prepared by methods known to those skilled in the art.

These groups of new compounds can be prepared in a manner known per se.

salts

Physiologically acceptable salts of the compounds according to the invention are prepared by methods known per se. These salts are novel compounds and are also an object of the invention. Metal salts can be prepared by treating a metal hydroxide with a compound of the invention. Examples of metal salts that can thus be prepared are those containing the metals Li, Na and K. A less soluble metal salt can be precipitated from a solution of a more soluble salt by adding a suitable metal compound. Acid salts can be prepared by treating a compound of the invention 80885 with an acid such as HCl, HBr, H 2 SO 4 or an organic sulfonic acid.

Pharmaceutical preparations

Pharmaceutical preparations of compounds of formula I.

O

A -

HhK \ I

k I> h2n

A

wherein A is (CH2) mCH- (CH2) nOH (Aa) or

Ra CH2-C = C - CH2OH (Ab)

Rb Rb wherein m is 1 or 2, n is 1 or 2, wherein m is 1 when n is 2 and m is 2 when n is 1, R4 is (CH2) pOH or CORE, R1 is hydroxy, p is 1 -4, and R b are independently selected from hydrogen and (CH 2) p OH, provided that at least one of R jj or R e is hydrogen, or R 1 and R b together form an additional carbon-carbon bond to form an alkyne and provided that when m is 2 and n is 1 and R 8 is (CH 2) p OH, then p is 2-4, or a physiologically acceptable salt, geometric or optical isomer thereof, together with a pharmaceutically acceptable carrier, form an embodiment of the process of the invention.

19 80885

The compounds of the invention may be administered systemically or topically, and in clinical practice are usually administered topically, orally, nasally, by injection, infusion or inhalation in the form of a pharmaceutical preparation containing the active ingredient in the form of the parent compound or optionally a pharmaceutically acceptable compound thereof. in the form of a salt together with a pharmaceutically acceptable carrier, which may be a solid, semi-solid or liquid diluent or using an ingestible capsule, and these preparations also form an embodiment of the invention. These compounds can also be used without a carrier. Examples of pharmaceutical preparations include tablets, solutions, drops such as nasal drops, eye drops, preparations for external use such as ointments, gels, creams and suspensions, inhalable aerosols, nasal sprays, liposomes, etc. The amount of active ingredient is usually 0.01 to 99% by weight. or 0.1-99% by weight, e.g. between 0.5-20%, in preparations for injection and between 0.1 and 50% in preparations for oral administration.

20 80885

The preparations are preferably in unit dosage form. Furthermore, they are preferably prepared in sterilized form.

To prepare pharmaceutical preparations in unit dosage form for oral use containing a compound of the invention, the active ingredient may be mixed with a solid, powdered carrier, for example lactose, sucrose, sorbitol, mannitol, starch such as potato starch, corn starch or laminaria powder, laminaria, amylopectin or gelatin, in which case lubricants such as magnesium or calcium stearate, or Carbowa> P 2 or some other polyethylene glycol wax, may also be used and compressed to form tablets or granular cores. In preparing the granules, the core parts can be coated, for example, with a concentrated sugar solution which may contain gum arabic, talc and / or titanium dioxide, or alternatively using a film-forming agent dissolved in a volatile organic solvent or mixture of organic solvents. Dyes may be added to these coatings, for example to distinguish between different concentrations of active substance. In preparing soft gelatin capsules containing gelatin and, for example, glycerol and a plasticizer, or in the form of such sealed capsules, the active ingredient may be mixed with Carbowaj or a suitable oil such as sesame oil, olive oil or arrack oil. Hard gelatine capsules may contain granules of the active ingredient together with solid, powdered carriers such as lactose, sucrose, sorbitol, mannitol, starches (for example potato starch, corn starch or amylopectin), magnesium strain or gelatin may also be used and cellulose derivatives or gelatin .

21 80885 The use of several layers of active drug separated by slowly dissolving coatings gives long-acting tablets. Another way to prepare long-acting tablets is to divide the dose of active drug into granules with different thicknesses of coatings and to compress the granules into tablets together with a carrier. The active ingredient may also be added to slowly dissolving tablets made of, for example, fat and wax or evenly distributed in a tablet formed by an insoluble substance such as a physiologically inert plastic substance.

Liquid preparations for oral administration may take the form of elixirs, syrups or suspensions, for example solutions containing from about 0.1 to 20% by weight of active ingredient, sugar and a mixture of ethanol, water, glycerol and propylene glycol, and optionally aromatics, saccharin and / or or carboxymethylcellulose as dispersants.

Parenteral preparations for injection may contain an aqueous solution of the active substance or a physiologically acceptable salt thereof, preferably in a concentration of 0.05 to 10%, and optionally also a stabilizing agent and / or a buffering agent in aqueous solution. Dosage units of the solution may preferably be enclosed in ampoules.

For topical use, especially in the treatment of herpes virus infections of the skin, genitals and mouth and eyes, these preparations are suitably in the form of a solution, ointment, gel, suspension, ointment and the like. The amount of active substance can vary, for example, between 0.05 and 20% by weight of active substance. Such preparations for superficial use can be prepared in a known manner by mixing the active ingredient with known carriers such as isopropanol, glycerol, paraffin, stearyl alcohol, polyethylene glycol, polypropylene glycol and the like. A pharmaceutically acceptable carrier may also contain a known chemical absorption enhancer. Examples of such absorption enhancers are, for example, dimethylacetamide 22,80885 (U.S. Patent 3,472,931), trichloroethanol and trifluoroethanol (U.S. Patent 3,891,757), certain alcohols and mixtures thereof (GB Patent 1,001,949). It may also contain detergents to promote the penetration of the active substance into the skin.

The dosages in which the active ingredients are administered can vary within wide limits and depend on various factors such as the severity of the infection, the age of the patient, etc., and these can be adjusted individually. Possible ranges of amounts of the compounds of the invention that may be administered per day include from about 0.1 to 2000 mg, preferably from 1 to 2000 mg, for surface administration, from 50 to about 2000 mg or from about 100 to 1000 mg for oral administration, and from about 10 to 2000 mg. mg or about 50-500 mg for injection.

In the most severe cases, it is necessary to increase the doses 5- or always 10-fold. In less severe cases, it may be sufficient to use 500-1000 mg of the compound.

Pharmaceutical compositions containing the active ingredients may conveniently be formulated so as to provide dosages within these limits in either a single dose or in multiple doses.

Thus, in accordance with the invention, it has been found that the compounds of the formula I and their physiologically acceptable salts are useful for inhibiting the growth of herpes viruses. The compounds of the formula I and their physiologically tolerable salts are useful in the therapeutic and / or prophylactic treatment of viral infections.

A preferred aspect of the invention is the use of a compound of formula I, including a compound wherein R 'is CH-OH, d L · when m is 2 and n is 1, or a physiologically acceptable salt thereof, in the treatment of herpes virus infections.

<23 80 8 8 5

Reference Examples

The compound of the invention 9- [4-hydroxy-2- (hydroxymethyl) butyl] guanine O

II

h / V \ c i I i N 1 -N — CH2-CH CH2-CH2OH VSA 671

H2N I

ch2oh to the closest prior art compound to 9- (2,4-dihydroxybutyl) guanine in FI Patent 68054 in Example 12 thereof, having the formula 0 hn, r 1 '1 X - - N - CH 2 -CH-CH 2 -CH 2 OH VIS 715

H2N I

OH

in the following Comparative Examples 1-3.

Comparative Example 1 Inhibition of plaque caused by HSV-1 in blood cells

Inhibition of the virus strain HSV-1 C-42 by VSA 671 and VIS 715 was measured in a plaque reduction assay in "Vero cells" (green monkey kidney cells) as described in Ejercito et al., J. Gen. Gen. Virol. 2,357 (1968). IC50 values were measured by close-up response studies at six concentrations of each substance. Values are the average of at least two different experiments.

Compound_ICc; n (μΜ) VSA 671 0.2 VIS 715 64 24 80 8 85

The values indicate that only 0.2 μΜ of VSA 671 is required to achieve a 50% reduction in HSV-1 plaque formation, whereas VIS 715 is required to achieve a corresponding reduction in plaque formation of 64 μΜ. The difference is therefore 320 times.

Comparative Example 2

Inhibition of Varicella zoster virus (VZV) replication in HL cells The replication of several VZV virus strains in HL cells (human lung connective tissue hemocytes) in the presence of VSA 671 and VIS 715 was determined. The assay was performed by measuring the formation of late antigens by ELISA as described in B. Wahren et al., J. Virological Methods 6, 141-149 (1983).

Compound_IC5Q (μΜ) VSA 671 1.5-2.5 (4 isolates) VIS 715 300 (1 isolate) To achieve 50% inhibition of VZV, VSA 671 1.5-2.5 μΜ is required, whereas VIS 715 is required as much as 300 μΜ. Such a high concentration indicates that VIS 715 is not a significant inhibitor of VZV formation.

Comparative Example 3

Affinity of VSA 671 and VIS 715 for Herpesvirus-Dused Thymidine Kinase Inhibition constants (K i) of VSA 671 and VIS 715 were determined to show the affinity of the compounds for HSV-1 thymidine kinase. The method was guided by A. Larsson et al., Antimicrob. Agents Chemother. 23, 664 (1983).

25 80 8 8 5

Compound__ VSA 671 0.6 VIS 715 19

The results show that VSA 671 has a much higher affinity for HSV-1-thymidine kinase than VIS 715. Such a high affinity promotes antiviral activity under conditions where the compounds have to compete with thymidine. The affinity of VSA 671 is close to that of the natural thymidine substrate, as its Km is 0.42 μΜ. The higher the K - ^ value, the lower the affinity of thymidine kinase.

26 80885

EXAMPLES

The following examples illustrate the preparation of the compounds of the invention.

Example 1. 9- / 4-Hydroxy-2- (hydroxymethyl) butyl / quanine (Method C) V I 671

H 2 N

CHoCHCH_CHo0H Z | z z ch2oh

A suspension of sodium borohydride (135 mg; 3.57 mmol) and dried lithium bromide (220 mg; 2.53 mmol) in 4 mL of dried and distilled diglyme was stirred at room temperature for 1/2 h.

After addition of dimethyl 2- (2-amino-6-chloropurin-9-ylmethyl) succinate (215 mg; 0.656 mmol) and 3 drops of trimethyl borate, the mixture was stirred at room temperature overnight and then kept at 100 ° C. 5 h. The solvent was evaporated in vacuo and the residue was dissolved in 10 ml of 70% formic acid in water and kept at 50 ° C overnight. The mixture was evaporated to dryness in vacuo, dissolved in water, neutralized with sodium bicarbonate and evaporated in vacuo. Chromatography (silica gel, ethyl acetate + methanol + water 4 + 3 + 1 parts by volume) and recrystallization from water gave 99 mg of 9- / 4-hydroxy-2- (hydroxymethyl) -butyl] guanine. The compound was purified by preparative HPLC (Porasil C 18 g reverse phase column, methanol + water 20 + 80) to give the pure product after recrystallization from water. Sp. 235 ° C.

UV spectrum, λ makg (nm): 0.01 M HCl 253 (277) H 2 O (pH 7) 252 (270 infl.) 0.01 M NaOH 268 (256 infl.) 27 80885 1 H NMR (DMSO-d 6) : δ 1.43 (m, 2H) CH 2 CH 2 OH; 2.09 (m, 1 H) CH; 3.34 (d, 2H) CHCH-OH; 3.47 (t, 2H) CH 2 <JH 2 OH; 3.96 (ABX system 2H) NCH 2; 6.25 (broad s, 2H) NH 2; 7.59 Ts, 1H) Hg, · 10.4 (broad s, 1H) NH.

The starting compound, dimethyl 2- (2-amino-6-chloropurin-9-ylmethyl) zincinate, was prepared as follows: (Method B)

Cl

1 jTS

2 I

CH2CHCH2COOCH3 C00CH3

A mixture of 2-amino-6-chloropurine (4.07 g; 0.024 mol), dimethyl itaconate (5.00 g; 0.032 mol) and sodium hydride (55% in oil; 0.2 g) in 50 ml: in dry dimethylformamide, was stirred at room temperature for 3 days. About 50 mL of water was then added and the mixture was washed with n-hexane (2 x 50 mL) and then extracted with 2 x 50 mL of dichloromethane. The combined CH 2 Cl 2 extracts were washed with 2 x 20 mL of water, dried over magnesium sulfate and evaporated in vacuo. Treatment with ether and drying gave a white crystalline product. Chromatography (silica gel, chloroform + methanol 15 + 1) gave 5.54 g (71%) and recrystallization (MeOH-H 2 O) 5.15 g (66.1%) of dimethyl-2- (2-amino-6-chloropurine). -9-ylmethyl) succinate.

UV spectrum in EtOH, λ max (nm): 310 (247) 1 H-NMR (CDCl 3) δ 2.67 (dd, 2H) CH 2 CO 2; 3.46 (m, 1 H) CH; 3.70 (2s, 2x3H) OCH3; 4.42 (ABX sequ. Jgem = 14 Hz, 2H) NCH 2; 5.35 (broad s, 2H) NH 2; 7.79 (s, 1H) Hg.

Example 2. 9- / 4-Hydroxy-3- (hydroxymethyl) butyl] guanine (Method A) 28 80 8 85 ° V VIS 873

H N

H2N1 ^ -CH2OH

CH2-CH2-ch

1 λ CH2OH

A mixture of 2-amino-6-chloropurine (1.275 g; 7.5 mmol) and potassium carbonate (0.69 g; 5 mmol) in dry dimethylformamide (30 mL) was sonicated for 15 min. benzyl 2-benzyloxymethyl-4-bromobutyl ether (1.82 g; 5 mmol) and the mixture was then allowed to stand in sunlight for 4 h and then stirred overnight. After evaporation in vacuo, the residue was extracted with 3 x 50 ml of chloroform and the extracts were evaporated to dryness in vacuo. Chromatography (silica gel, chloroform + methanol 20 + 1 by volume) gave 0.58 g (26%) of pure compound, 4- (2-amino-6-chloropurin-9-yl) -2-benzyloxymethyl-butyl-benzyl ether. . A small sample of this compound was recrystallized from toluene. Sp. 108.5 to 109.5 ° C.

1 H NMR (CDCl 3): δ 1.96 (m, 3H) N-C-CH 2 CH; 3.48 (ABX system 4H) CH (CH 2 O) 2; 4.13 (t, 2H) NCH 2; 4.46 (sT 4H) OCH 2 Ph; 5.49 (s, 2H) NH 2; 7.30 (m, 10H) phenyl; 7.69 (s, 1 H) Hg.

Cl πΛ ^ Ν H2N \ I ch2och2- (O) CH2-CH2-CH '-' ch2och2 -

Unprotecting

A solution of 4- (2-amino-6-chloropurin-9-yl) -2-benzyloxymethylbutylbenzyl ether (0.904 g; 2 mmol) in 25 mL of 80% aqueous formic acid was maintained at 100 ° C. 1 h, palladium catalyst (10% on carbon; 29 80885 0.40 g) was added and the mixture was hydrogenated at room temperature and atmospheric pressure with vigorous stirring for 45 min. After evaporation to dryness in vacuo, the residue is heated to boiling point together with 50 ml of water, 1 ml of concentrated aqueous ammonia solution are added, the solution is gently boiled briefly and then evaporated to dryness. Recrystallization from water gave 0.23 g (45%) of 9- [4'-hydroxy-3- (hydroxymethyl) butyl] guanine. Sp. 252-262 ° C (decomposes, slow heating) or 264-265 ° C (rapid capillary change).

1 H NMR (DMSO-d 6): 61.51 (m, 1H) CH; 1.76 (q, 2H) N-C-CH 2; 3.42 (ΔBX system 4H) 2CH 2 O; 4.02 (t, 2H) NCH 2; 6.29 (broad s, 2H) NH 2; 7.64 (s, 1 H) H 8; 10.3 (broad s, 1H) NH.

UV, max (nm): ethanol 255 ε12300 (~ 270 infl.) 1 M HCl 253 C11300 279 1 M NaOH 268 (257 infl.) The starting compound, benzyl-2-benzyloxymethyl-4-bromobutyl ether, was prepared as follows: a ) Diethyl (2,2-dimethoxyethyl) malonate CH 3 O / COOCH 2 -CH 3

^ CH-CH2-CH

CH-JO "COOCH2-CH3

Diethyl malonate (168.18 g; 1.05 moles) was added to a solution of sodium ethoxide (23.0 g; 1.00 moles of sodium) in 500 ml of dry ethanol at 50 ° C over 45 min. Bromoacetaldehyde dimethyl acetal (74.2 g; 1.06 mol) was then added at the same temperature over 45 minutes and the mixture was heated at reflux overnight. The ethanol was evaporated in vacuo and the residue was dissolved in 200 ml of water and extracted with 3 x 300 ml of ether. The extracts were dried (MgSO 4), evaporated and distilled in vacuo to give 115.0 g (46%) of the title compound. Kp. 97-101 ° C (0.6 mm Hg).

80885 1 H NMR (CDCl 3): δ 1.30 (t, J = 7Hz, 6H) 2 C-CH; 2.20 (dd, J = 5 Hz and 7 Hz, 2H) CH-CH 2; 3.35 (s, 6H) 2 OCH 3; 3.44 (t, 1H) COCHCO; 4.20 (q, J = 7 Hz, 4 H) 2 OCH 2; 4.42 (t, J = 5 Hz, 1H) O-CH-O.

b) 4,4-dimethoxy-2- (hydroxymethyl) butanol

CH-, 0 CHoOH

3 \ ./ 2

CH-CH "-CH

z '\ CH30 ch2oh

Diethyl (2,2-dimethoxyethyl) malonate (115.0 g; 0.46 mol) was added to a suspension of lithium aluminum hydride (38.0 g; 1.00 mol) in 400 ml of dry ether at such a rate that it was possible to maintain careful reflux. After stirring for 15 minutes, 40 ml of water, 40 ml of 15% aqueous NaOH solution and 120 ml of water were added, and stirring was therefore continued until a white suspension was obtained. The inorganic salts were filtered off and extracted with several 200 ml portions of tetrahydrofuran (THF). The combined organic extracts were evaporated and distilled twice in vacuo to give 50.23 g (66.5%) of the title compound. Kp. 116-125 ° C (0.02 mm Hg).

1 H NMR (CDCl 3): δ 1.6-2.1 (m, 3H) CH 2 CH; 3.35 (s, 6H) 2 OCH 3; 3.67 (d, 4H) 2 CH 2 O; 4.48 (t, J = 5 Hz, 1H) O-CH-O.

c) Benzyl 2- (benzyloxymethyl) -4,4-dimethoxybutyl ether ch3o ch2och2 - (5)

N''S 'CH-CH - CH

X \ / ΤΛ CH30 CH2OCH2 - \ Q /

Sodium hydride (55% in oil; 32.72 g; 0.75 mol) was washed twice with n-hexane and suspended in 200 ml of THF. 4,4-Dimethoxy-2- (hydroxymethyl) butanol (49.26 g; 0.3 mol) was then added dropwise to maintain a slight evolution of hydrogen gas. Benzyl chloride (94.94 g; 0.45 mol) was added over 31 minutes and the mixture was heated at reflux overnight, cooled, diluted with 50 ml of ethanol and evaporated in vacuo. The residue was suspended in about 200 ml of water and extracted with 3 x 100 ml of ether. The combined organic extracts were dried (MgSO 4) and distilled in vacuo. The title compound (94.34 g; 91%) was obtained at 197-200 ° C (0.01 mm Hg).

1 H NMR (CDCl 3); Δ 1.67 (indistinct t, 2H) CHCH 2 CH; 2.02 (m, 1 H) CH; 3.23 (s, 6H) 2 OCH 3; 3.46 (d, 4H) 2 CH 2 O; 4.43 (s, 4H) 2 OCH 2 Ph; 4.43 (t, 1H) O-CH-O; 7.27 (broad s, 10H), phenyl.

d) 4-Benzyloxy-3-benzyloxymethyl-butan-1-ol-ch2och2 - (c)

IIOCH2-CH 2 CH

\ ch2och2 - (o)

A mixture of benzyl 2-benzyloxymethyl-4,4-dimethoxybutyl ether (51.62 g; 0.15 mol), acetic acid (70 ml), water (30 ml) and THF (100 ml) was heated to reflux for 2 hours. h and then evaporated in vacuo. The residue was dissolved in ether and the solution was washed with aqueous potassium carbonate solution and water, dried (MgSO 4) and evaporated in vacuo. The residue of crude 4-benzyloxy-3-benzyloxymethylbutanal (quantitative yield) was dissolved in 100 ml of THF and added to THF over 30 minutes. to a suspension of lithium aluminum hydride (7.6 g; 0.20 mol) in 100 ml of THF.

8 ml of water, 8 ml of 15% aqueous NaOH solution and 24 ml of water were then added, and the mixture was stirred until it turned white to give a cloudy suspension. The inorganic salts were filtered off and washed with several portions of THF. The combined extracts were evaporated in vacuo to give 41.93 g (93%) of the title compound.

1 H NMR (CDCl 3): δ 1.66 (q, 2H) CH 2 CH 2 O; 2.05 (m, 1 H) CH; 3.0 (t, 1H) OH; 3.46 (d, 4H) CH (CH 2 O 2; 3.64 (q, 2H) CH 2 OH; 4.47 (s, 4H) 20 CH 2 Ph; 7.30 (m, 10H) phenyl).

32 80885 e) Benzyl 2-benzyloxymethyl-4-bromobutyl ether ch2och2-

BrCH - CH ~ -CH

^ ch2och2 -

A solution of 4-benzyloxy-3-benzyloxymethylbutan-1-ol (41.75 g; 139 mmol) in 150 mL of dichloromethane was cooled with an ice-water bath. Triphenylphosphine (42.0 g; 160 mmol) was then added with stirring, followed by N-bromosuccinimide (26.0 g; 146 mmol) portionwise over 15 minutes. After standing for 1 h at room temperature, ether (400 ml) was added and the precipitated triphenylphosphine oxide was filtered off.

The solution was concentrated in vacuo to a small volume and more ether was added to the extract to give precipitated triphenylphosphine oxide. Finally, this extraction-precipitation step was repeated using light petroleum to give, after evaporation, 33.55 g (66%) of the crude title compound.

1 H NMR (CDCl 3): 1.98 (q, 2H) Br-C-CH 2; 2.15 (m, 1 H) CH; 3.45 (t, 2H) BrCH 2; 3.47 (m, 4H) CH (CH 2 O) 2; 4.47 (s, 4H) 2 OCH 2 Ph; 7.30 (m, 10H) phenyl.

Example 3. 9- / T-hydroxy-2- (hydroxymethyl) butyl / guanine (Method C) 0 HNuX> VSA671

h_n '"^ n 2 I

CH 2 CH CH2CH2OH

CH 2 OH

Dimethyl 2- (2-amino-6-chloropurin-9-ylmethyl) succinate (8.00 g; 0.0244 mol) was dissolved in 200 ml of tert-butanol at 50 ° C, then lithium borohydride 33 80885 (2 , 66 g; 0.122 mol) and the mixture was stirred for 1 h. Then 100 ml of water were added and stirring was continued overnight, the inorganic salts were filtered off and the solution was evaporated to dryness in vacuo. The residue was found to contain 78 mol% of 2-amino-6-tert-butoxy-9- / 4-hydroxy-2- (hydroxymethyl) butyl-7-purine together with the 6-chloro compound.

1 H NMR (DMSO-d 6): δ 1.10 (s, 9H) C (CH 3) 3; 1.38 (m, 2H) CH 2 CH 2 OH; 2.14 (m, 1 H) CH; 3.34 (d, 2H) CHCH 3 OH; 3.45 (indistinct t, 2H) CH 2 CH 2 OH; 4.06 (t, ABX system 2H) NCH 2; ~ 4.5 (very broad s) OH; 6.77 (s, 2H) NH 2; 8.08 (s, 1H) HQ.

13 C NMR (DMSO-d 6): 631.41, C (CH 3) 3; 31-92, CH2CH2OH; 37.88, CH; 45.01, NCH 2; 58.72 and 61.3, 2CH2OH; 66.65, C (CH 3) 3; 123.59, C5; 143.91, C8; 149.53, C6; 154.59, C4; 159.87, C2.

OC (CH3) 3

fifV

h2n,

CH2CHCH2CH2OH

CH20H

Unprotecting

A solution of crude 2-amino-6-tert-butoxy-9-t-hydroxy-2- (hydroxymethyl) butyl / purine (436 mg; 1.41 mmol) in 6 mL of 50% aqueous formic acid , kept at 100 ° C for 3 h and then evaporated to dryness in vacuo, more water was added and the mixture was lyophilized. The residue was kept at 100 ° C for 2 h together with 10 ml of 2M aqueous ammonia solution. The ammonia was removed with a stream of nitrogen gas, the solution was heated to boiling point, a small amount of solid was removed and the filtrate was cooled in a condenser to give 208 mg of crystalline 9- / 4-hydroxy-2- (hydroxymethyl) butyl / guanine.

13 C NMR (DMSO-d 6): δ 31.80, CH 2 CH 2 OH; 38.17, CH; 44.35, NCH 2; 58.80 and 60.94, 2CH2OH; 116.64, C5; 138.05, C8; 151.57, C4; 153.57, C2; 156.92, C6.

34 80885

Example 4. 4- (2-Amino-1/6-dihydro-6-oxopurin-9-yl) -2-hydroxymethylbutyric acid (Method A) -R, N '

2 I

CH2CH2CHCH2OH

COOH

2-amino-6-chloropurine (3.37 g; 19.9 mmol), 4-bromo-2-hydroxymethylbutyric acid ethyl ester (4.49 g; 19.9 mmol) and anhydrous potassium carbonate (2.75 g; 19.9 mmol) was mixed with 50 ml of dimethylformamide and the mixture was stirred at room temperature for 92 hours. The mixture was then filtered and the filtrate was evaporated. The residue was extracted with 3 x 50 ml of chloroform, and the combined extracts were evaporated. Chromatography (silica gel, chloroform-methanol 10: 1) and two recrystallizations from water gave 0.995 g of 4- (2-amino-6-chloro-purin-9-yl) -2-hydroxymethyl-butyric acid ethyl ester.

13 C NMR (CDCl 3): ppm 173.56, 159.40, 153.81, 151.16, 142.38, 124.89, 62.61, 61.11, 45.08, 41.91, 28.46 and 14.16.

Cl cYs h2n '^ n I *'

CH2CH2CHCH20H

COOCH2CH3

Deprotection 4- (2-Amino-6-chloro-purin-9-yl) -2-hydroxymethyl-butyric acid ethyl ester (0.80 g) in 50 ml of 1M hydrochloric acid was heated at reflux for 3 h and then evaporated in vacuo. Re-evaporated with 3 x 10 ml of water and the residue was dissolved in 5 ml of water. The pH was then adjusted to 6-7 with solid sodium bicarbonate. The resulting solution was filtered and the pH was adjusted to 4 with acetic acid. After cooling to 0 ° C, the precipitate was filtered off and washed with water. Recrystallization from water gave 0.43 g of 4- (2-amino-1,6-dihydro-6-oxopurin-9-yl) -2-hydroxymethylbutyric acid.

13 C NMR (DMSO-d 6): ppm 174.80 (C = O), 157.09 (C6), 153.64 (C2), 151.38 (C4), 137.49 (C8), 116.84 ( C5), 62.15 (CH2OH), 45.59 (CH), 41.33 (NCH2) and 28.66 (N-CH2-CH2).

The starting compound, 4-bromo-2-hydroxymethylbutyric acid ethyl ester, was prepared as follows:

BrCH2CH2CHCH20H

cooch2ch3 2-hydroxymethylbutyrolactone / G. Claesson et al., Arkiv för kerni 2J3, 173 (1967 | 7 (3.0 g)) was dissolved in 20 ml of ethanol and the solution was saturated with hydrogen bromine at 0 [deg.] C. After standing for 0.5 h at 0 [deg.] C., the solvent was evaporated under reduced pressure. The mixture was then extracted several times with diethyl ether and the combined ether extracts were washed with saturated aqueous sodium chloride solution and dried over anhydrous sodium sulfate. Evaporation of the solvent gave 4.59 g of 4-bromo-2-hydroxymethyl-butyric acid ethyl ether.

13 C NMR (CDCl 3): ppm 174.14, 62.54, 61.08, 46.02, 31.45, 38.89 and 14.26.

Example 5. 9- [4-Hydroxy-2- (hydroxymethyl) butyl] guanine hydrochloride 9- [4-hydroxy-2- (hydroxymethyl) butyl] guanine (46 mg; Example 1) was dissolved in 0.4 ml of 1 M hydrochloric acid. the aqueous solution and the solution were lyophilized (0.1 mm Hg) overnight. The residue (53 mg; quantitative yield 3680885) was dissolved in a few drops of warm ethanol and precipitated using several volumes of tetrahydrofuran to give a white crystalline product. Sp. about 85 ° C (d).

1 H NMR (D 2 O): δ 1.61 (m, 2H) CH 2 CH 2 OH; 1.87 (m, 1 H); 2.31 (septet, 1H) CH; 3.57 (d, 2H) CHCH 2 OH; 3.72 (unclear q) CH 2 CH 2 OH; 4.32 (m, 2H) NCH 2; 4.90 (s) HDO; 8.96 (s, 1H) Hg.

Example 6. 9- [4-Hydroxy-3- (hydroxymethyl) -2-butenyl] guanine (Method A) -0 HN | VSA 668 h2n N 1 ^ .ch2oh

CH "CH = C

z ^ CH2OH

Crude 1-benzyloxy-2-benzyloxymethylbut-3-en-2-ol (1.42 g; 3.16 mmol) in 10 mL of CH 2 Cl 2 was added to 0.71 g of thionyl chloride (6 mmol) and 2 drops of DMF in 10 mL in CH 2 Cl 2 and the solution was stirred for 0.5 h and evaporated in vacuo. Toluene was then added and the mixture evaporated in vacuo to give crude 1-benzyloxy-2-benzyloxymethyl-4-chloro-but-2-ene.

A mixture of 1.61 g of 2-amino-6-chloropurine (9.5 mmol) and 1.31 g of finely ground anhydrous K 2 CO 3 (9.5 mmol) in 50 ml of dry dimethylformamide (DMF) was kept at 100 ° C for 5 min and cooled. At 10 ° C, crude 1-benzyloxy-2-benzyloxymethyl-4-chlorobut-2-ene in 25 mL of dry DMF was added with stirring over 4 h.

After 2 days at room temperature, the mixture was evaporated in vacuo, the residue was extracted with 2 x 25 ml of chloroform and the extract was evaporated to dryness. Chromatography (silica gel CHCl 3 + MeOH 19 + 1) gave 0.25 g (18%) of 4- (2-amino-6-chloropurin-9-yl) -1-benzyloxy-2-benzyloxy-3780885 methyl-2 -butene (TLC on silica gel, CHCl 3 + MeOH 13 + 1 vol, Rf 0.62).

NMR (CHCl 3 -CD 3 OD): 64.05 and 4.19 (2s, 2x2H) = C (CH 2 O) 2; 4.49 and 4.55 (2s, 2x2H) benzyl CH 2; 4.70 (d, 2H) N-CH 2; 5.85 (t, 1H) CH =; 6.2 (broad s, 2H) NH 2; 7.3 (broad s, 10H) phenyl; 7.49 (s, 1 H) Hg.

Cl «Vs H2N I ^ CH2OCH2C6H5 ch2ch = c.

1 ^ ch-OCh ^ c, ^ 2 2 6 5

Unprotecting

A solution of 4- (2-amino-6-chloropurin-9-yl) -1-benzyloxy-2-benzyloxymethyl-2-butene (148 mg; 0.329 mmol), 1.8 mL of trifluoroacetic acid and 0.2 After cooling, the mixture was diluted with 2 ml of water, heated to reflux for 1 minute, evaporated to dryness in vacuo, dissolved in water and re-evaporated by chromatography (silica gel, ethyl acetate + methanol + methanol). 70 + 20 + 1 parts by volume) gave 34 mg (41%) of 9- [4-hydroxy-3- (hydroxymethyl) -2-butenyl] guanine.

1 H NMR (DMSO-d 6): 63.96 and 4.10 (2d, 2x2H) = C (CH 2 O) 2; 4.68 (d, 2H) N-CH 2; 4.83 (2t, 2H) 2 OH; 5.60 (t, 1H) CH =; 6.50 (broad s, 2H) NH 2; 7.66 (s, 1H) Hg, · 10.6 (broad s, 1H) NH.

The starting compound, 1-benzyloxy-2-benzyloxymethyl-but-en-2-ol, was prepared as follows: HO 2 CH 2 OCH 2 C 6 H 5 CH 2 = CH CH 2 OCH 2 C 6 H 5

A solution of 4.0 g of vinyl bromide (37.4 mmol) in 8 mL of dry tetrahydrofuran (THF) was added to 0.94 g of Mg (38.7 mmol) in 3 mL of THF under N 2 at about 60 ° C and the mixture was heated to reflux (CO 2 -ethanol condenser) for 1 h.

38 8 0 8 8 5

A solution of 7.4 g (27.4 mmol) of 1,3-bis (benzyloxy) -propan-2-one (Araki Y. et al. J. Chem. Soc. Perkin Trans. 1, 1981, 19) in THF, was added dropwise at 10 ° C and the mixture was stirred at ambient temperature for 2 h. Then 8 ml of a 20% aqueous NH 4 Cl solution were added and the organic phase was evaporated, dissolved in ether, dried over MgSO 4 and evaporated in vacuo to give 7.90 g of 1-benzyloxy-2-benzyloxymethyl-but-en-2-ol as a crude oil.

1 H NMR (CDCl 3): 52.67 (s, 1H) OH; 3.50 (s, 4H) C (CH 2 O) 2; 4.55 (s, 4H) 2 OCH 2 Ph; 5.0-6.3 (m, 3H) CH 2 = CH; 7.32 (broad s, 10H) phenyl.

Example 7. cis-9- (4-hydroxy-2-butenyl) guanine (Method B) 0 H Ν 'Y \ I 11 / VSA 151

· HjnNt '

CH. .CH.OH

2 \ y 2 c = c X \

H H

2-Amino-6-chloropurine (3.4 g), butadiene monoepoxide (1.65 g) and tetrakis (triphenylphosphine) palladium (0) (1.0 g) in dry dimethylformamide (75 ml) were stirred under nitrogen and heated at 60 ° C. ° C for 45 minutes.

The solvent was evaporated in vacuo (0.1 mm Hg) and the residue was triturated with hot chloroform (4 x 50 ml). The combined chloroform solutions were concentrated (to a volume of 25 mL) and diluted with ether (100 mL). The resinous precipitate was collected, dried and triturated with hot 1,2-dimethoxyethane (3 x 50 mL). The combined solutions were evaporated in vacuo to 3380885 (0.1 mm Hg) to give a crude mixture (4 g) which was separated on a silica gel column (500 g), eluted with chloroform + methanol 10 + 1 (800 ml) and then chloroform + methanol 8 1. Fractions of about 15 ml were collected and the compound cis-4- (2-amino-6-chloropurin-9-yl) -2-buten-1-ol was obtained as a smaller product (0.2 g) from fractions 74-78. Sp. 160-165 ° C.

UV spectrum, λ (nm): 0.1 M HCl 241, 315 0.1 M NaOH 246, 308 1 H NMR (DMSO-d 6): δ 4.20 (t, J 5.4 Hz, 2H) CH 2 O; 4.76 (d, J 6.8

Hz, 2H) NCH 2; 5.54-5.83 (m, 2H) CH = CH 2. 6.69 (s, 2H) NH 2; 8.06 (s, 1H) Hg.

Cl

N ^ V ^ V

H „N N I H H

+ I i I

CH2-C = C-CH2OH

Deprotection cis-4- (2-amino-6-chloropurin-9-yl) -2-buten-1-ol (50 mg) in formic acid water 7 + 3 (5 ml) was heated at 45 ° C under nitrogen for 20 hours. The solvent was evaporated in vacuo and the residue was dried. Water (0.5 ml) was then added and the solution was neutralized with aqueous ammonia. The precipitate was filtered, washed with water, ethanol and ether and dried to give cis-9- (4-hydroxy-2-butenyl) guanine (40 mg).

1 H NMR (DMSO-d 6): 64.15 (indistinct t, 2H) CH 2 O; 4.65-4.90 (ABX system 2H) NCH 2; 5.75 (m, 2H) CH = CH; 6.52 (broad s, 2H) NH 2; 7.66 (s, 1 H) Hg.

Example 8. 9- (4-Hydroxy-2-butynyl) guanine (Method A) 0

Jk / N.

I) VSB 631

H with S ^^

CH2C = CCH2OH

80885 N-Bromosuccinimide (1.78 g; 10 mmol) was added portionwise to a stirred suspension of 2-butyn-1,4-diol (2.50 g; 30 mmol) and triphenylphosphine (2.62 g; 10 mmol). In 20 ml of dichloromethane at room temperature.

After 1 h, the unreacted butyndiol was removed by filtration. The solution was evaporated in vacuo to a small volume, dissolved in 20 ml of dry dimethylformamide (DMF) and added to a suspension of 2-amino-6-chloropurine (1.70 g; 10 mmol) and finely ground anhydrous K 2 CO 2 (2.07 g). 15 mmol) in 75 mL of dry DMF and stirred at 0 ° C. Stirring was continued at room temperature for 3 days. The mixture was then evaporated to dryness in vacuo. Chromatography (silica gel, CHCl 3 + MeOH 7 + 1) gave 467 mg (20% after recrystallization from water to remove 2-butyn-1,4-diol) 4- (2-amino-6-chloropurin-9-yl) -2-butyn-1-ol (TLC on silica gel, CHCl 3 + MeOH 5 + 1, 0.57).

1 H NMR (DMSO-d 6): δ 4.10 (dt, 2H) CH 2 O; 4.96 (t, J = 2.0 Hz, 2H) NCH 2; 5.12 (t, J = 5.9 Hz, 1 H) OH; 6.89 (broad s, 2H) NH 2; 8.17 (s, 1H) Hg.

13 C NMR (DMSO-d 6): δ 32.79 (NCH 2); 49.07 (CH 2 O); 77.80 and 85.19 (CSC); 123.43 (C5); 142.43 (C8); 149.82 (C6); 153.69 (C4); 160.01 (C2).

Cl H2N '' ch2c Ξ cch2oh

Unprotecting

A solution of 4- (2-amino-6-chloropurin-9-yl) -2-butyn-1-ol (140 mg; 0.59 mmol) in 2 mL of 50% aqueous formic acid was maintained at 80 ° C. 6 h and then evaporated to dryness in vacuo. The residue was dissolved in 2 ml of water + concentrated aqueous ammonia 1 + 1 and the solution was kept at 80 ° C for 1 h and then evaporated to dryness in vacuo.

4i 80 8 85

Recrystallization from water gave 90 mg (70%) of 9- (4-hydroxy-2-butynyl) guanine.

1 H NMR (DMSO-d 6): δ 4.09 (m, 2H) CH 2 O; 4.84 (t, J = 2.0 Hz, 2H) NCH 2; 5.11 (t, J = 6.0 Hz, 1 H) OH; 6.44 (broad s, 2H) NH 2; 7.73 (s, 1 H) Hg.

Biological experiments

The inhibitory effect of the compounds of the invention against the herpes virus was tested using the methods described below. The toxicity of the compounds used to basal cell function was also tested.

The following is the entry VSA 671 for 9- [4-hydroxy-2- (hydroxymethyl) butyl] guanine (base), the entry 9- [4-hydroxy-3- (hydroxymethyl) butyl] guanine (base) is VIS 873 , for cis-9- (4-hydroxy-2-butenyl) guanine (base) the designation VSA 151 and for the compound 9- [4-hydroxy-3- (hydroxymethyl) -2-butenyl] guanine (base) the designation VSA 668, Herpes simplex type 1 virus is designated HSV-1, Herpes simplex type 2 virus is designated HSV-2, and Varicella zoster virus is designated VZV.

42 80885 I. Inhibition of virus replication in cellular structures The inhibitory effect of HSV-1 virus (strain C42) using VSA 671, VIS 873, VSA 151 and VSA 668 was measured in a plaque reduction experiment using Vero (green monkey kidney) cells and using HEL- cells (human lung embryo) as described by Ejercito et al., J.Gen.Virol. 2.357 (1968). The effect on VZV (multiple strains) proliferation in HL (human lung tissue) cells by VSA 671 was determined by measuring late antigen formation using ELISA as described by B. Wahren et al., J. Virological Methods 6, 141-149 ( 1983). The results are shown in Table 1.

table 1

Inhibition of herpes virus replication by VSA 671, VIS 873, VSA 151 and VSA 668 _HSV-1_ VZV_

Compound in Vero cells in HEL cells In HL cells inhibition (%) / conc. ID, - (/ um) ID50 ^ / um ^ ___ (/ um) _ ___ _ VSA 671> 90/1 0.2 1.3-2.5 :: VIS 873> 90/1 0.4 VSA 151> 90/10 0.6 VSA 668> 90/25 II. Affinity of VSA 671, VIS 873, VSA 151 and VSA 668 for herpesvirus-induced thymidine kinase

Inhibition constants (K) showing affinity for HSV-1-induced thymidine kinase were determined using the method of A. Larsson et al., Antimicrob. Agents Chemother.

23, 664 (1983). The inhibition constants are shown in Table 2.

Table 2

Inhibition constants of VSA 671, VIS 873, VSA 151 and VSA 668 against HSV-1 virus-induced thymidine kinase

Compound K.

i _ (, um) VSA 671 0.6 VIS 873 1.5 VSA 151 7 VSA 668 25 «80885

The high affinity for viral thymidine kinase is an advantage for antiviral activity under conditions where these compounds have to compete with thymidine. The affinity of VSA 671 is close to that of the natural substrate, thymidine (Km 0.42 μm).

III. Cellular toxicity

The cellular toxicity of VSA 671, VIS 873, VSA 151 and VSA 668 was tested in human embryonic cells as described by K. Stenberg (Biochemical Pharmacology 30, 1005-1008 (1980). All of these compounds had low cellular toxicity (Table 3) at concentrations , which inhibit herpesvirus replication by more than 90% (see Table 1).

Table 3

Cellular toxicity of VSA 671, VIS 873, VSA 151 and VSA 668 as percentage reduction in cell growth (Flow 1000) after 48 h of incubation_ Concentration Compound Decrease in cell growth (μm) _ _ %_ 200 VSA 671 18 200 VIS 873 7 200 VSA 151 0 200 VSA 668 0 IV. Animal experiments a) Experiments on skin herpes virus infection in guinea pigs were performed as previously described by S. Alenius and B. Öberg / Archives of Virology 58, 277-288 (1978). The experiments showed that VIS 873 has a therapeutic effect. for herpes virus skin infections when administered superficially as described in Table 4. 30 ul of solution or solvent was superficially administered three times a day for 4 days starting 24 h after implantation.

44 80885

Table 4

Therapeutic effect on herpes virus infection on the skin

Concentration of VIS 873%% reduction in cumulative value in dimethyl sulfoxide compared to placebo __% _ ___ 2.5 36 b) Treatment of "systemic" HSV-1 or HSV-2 infection in mice using VSA 671 or VIS 873 i.p. was performed as reported by A. Larsson et al., Antimicrob. Agents Chemother. 2J3, 664-670 (1983).

5

Mice were infected using 10 pfu (plaque forming units) of HSV-1 (C42) or 10 pfu of HSV-2 (strain 91075) i.p. and treatment was started 1 h after infection. The treatment consisted of two daily i.p. dose of each nucleoside analog dissolved in phosphate buffered saline, 5 days. Cumulative mortality (treated vs. untreated animals) was determined during the experiment, i. Within 21 days. The results are shown in Table 5.

Table 5

Antiviral activity of VIS 873 and VSA 671 on systemic (i.p.) HSV-1 · and HSV-2 infections in mice

Compound Dose Virus Cumulative mortality (%) _ _ _ treated / untreated) VSA 671 25 mg / kg HSV-1 30/90 50 "" 30/90 25 HSV-2 40/90 50 "" 30/90 VIS 873 5 mg / kg HSV-1 30/80 10 "" 0/80 5 "HSV-2 70/90 10" "20/90 25" "0/90 50" "10/100 45 80885

Best Mode for Carrying Out the Invention

Of the compounds of formula I according to the invention, the compounds 9- / T-hydroxy-3- (hydroxymethyl) butylguanine, 9- / 4-hydroxy- (2-hydroxymethyl) butyl] guanine and cis-9- (4-hydroxy-2- butenyl) guanine and their use in the treatment of herpes virus infections is currently the best known embodiment. The compound 9- / 4 "-hydroxy-2- (hydroxymethyl) -butyl] guanine has been shown to have a very good effect against the replication of Varciella zoster virus.

Claims (12)

1 HN 8 I 2J 4L / H9N NN 3 19 A in which Formula A on (CH2) m-CH- (CH2) nOH (Aa) or R <k CHo-C = C-CHoOH (Ab) IIR ύ Rb in which formulas m are 1 or 2, n is 1 or 2, m is 1 where n is 2 and m is 2 where n is 1, Rq is (CH 2) pOH or COR 2, R 2 is hydroxy, p is 1-4 , R 2 and R 2 are independently selected from hydrogen and the compound (CH 2> pOH, provided that at least one of the groups R 2 and R 2 is hydrogen, or R 2 and R 2 together form a carbon-carbon addition bond such that an alkyne is formed, and further provided that when m is 2 and n is 1 and Rq is (CH2) pOH, then p is 2-4, or a physiologically suitable site, a geometric or optical isomer thereof, in the preparation of a compound of formula I, wherein A is Aa, characterized in that a compound of formula R2a xa '(ch2) m "CH" <CH2) n- ° R3a is condensed to the N-9 position in a guanine derivative of formula R6a xX> R4a-N NNII R5a Y after which any protecting groups of Xa Sr is a leaving group such as chlorine, bromine, iodine or OSO2R7a, wherein R7a is alkyl, fluorinated alkyl, arylalkyl or aryl, Y is hydrogen or a quaternary ammonium ion, R3a is hydrogen or protecting group such as acyl, alkoxycarbonyl, arylcarbonyl, silyl or SO 2 R 7a, R 2a is Ra as defined above or CN, COORga, CH (ORga) 2, CH (SRga) 2, NHCONHRga or (CH2) pORga, where p is defined as above , Rga is a substituted silyl or the group Rya as defined above, Rga is the group Rga as defined above, or Rga and Rga together with the group R3a form a "cross-linked group ie. a cyclic derivative such as a lactone, cyclic ester, ortho acid, acetal, ether or a silyl type compound, R4a and Rtja are the same as the group R3a above. R6a is a leaving group such as the group Xa, OH or OR defined above. Wherein R10a is an alkyl, arylalkyl, SO2R7a, substituted silyl, phosphoryl diester or phosphinothioyl, B. a compound of formula R14a CH2 = C - CH2 - Risa 54 80 885 is condensed to the N-9 position in a guanine derivative of formula R6a No. II R5a H and then any protecting groups are removed in which formulas R4a 'R5a and R6a are defined as in point A, R14a are CN, CONH2 or COORga, and Risa Sr COORga or CHaORga, wherein R8a and R9a s & as defined in point AC are derivatized or activated carboxylic acid groups in a compound of formula R6a NI >> κ48-νΑΝΛν / R12a R5a (CH2) m-CH- (CH2) n_i-COR11a are reduced, whereupon any protecting groups are removed, wherein formula m, n, R2a, R4a 'R5a and R6a are the same as definitions is, in point A, Rna Sr is the group Rga or the group (CH3) 2N = CHO-, and Ri2a Sr is the group R2a or the group (CH2) p_iCOR, where p is as defined in point AD a compound of formula R2b R7b Xb-CH2-C = C-CH2OR3b is condensed to the N-9 position in a guanine derivative of the formula 55 80 8 85 R6b / N. I II ^ JL I / R4h-N ^ N ^^ iT II R5b Y and then any protecting groups are removed, in which formulas Xb is a leaving group such as chlorine, bromine, iodine or OSC ^R5 b 'where Rgb is an alkyl, fluorinated alkyl, arylalkyl or aryl, Y is hydrogen or quaternary ammonium ion, R 2b is hydrogen or a protecting group such as acyl, alkoxycarbonyl, aryloxycarbonyl, silyl or SO2 Rgb, R2b is Rb or (CH2) pOR9b and R7b is R ' or (CH2) pORgb, where p is 1-4, and where Rgb is the group R3b as defined above, or Rgb and R3b together form a cross-linked group i.e. a cyclic derivative such as a carbonate ester, thiocarbonate ester, ortho acid or acetal compound, Rgb is a hydroxy or a leaving group such as chlorine, bromine, iodine, mercapto, alkylthio, SO2RQb or OR10b 'where R3 is an arylalkyl, substituted silyl, phosphoryl diester, phosphinothioyl or SO2 Rgb / R4b and R5b are the same as the group defined above R3b, E. A compound of the formula R2b R7b CH0 = C - C - CH2 O is condensed to the N-9 position in a guanine derivative of formula R6b II R5b H see 80885 whereupon any protecting groups are removed in which formulas R2b, R4b 'R5b' R6b and R7b are defined in point D, whereupon the obtained compound is optionally converted to a physiologically suitable salt or a geometric isomer. A process for the preparation of a therapeutically active compound of the formula 0 6.1 n Process according to Claim 1, characterized in that a compound is prepared having the formula 0 II / 11 HoN (CH2) m- CH- (CH2) nOH R4 in which formula m is 1 or 2 and n is 1 or 2, wherein m is 1, where n is 2 and m is 2, where n is 1, Rg is (CH2) pOH or CORg, where Rg is hydroxy, and p is 1-4 under the further condition that m is 2 and n is 1 and R 9 is (CH 2) pOH, s is p 2-4, and its physiologically suitable salts and optical isomers. 3. A process according to claim 1, characterized in that a compound is prepared having the formula O II-III ch2 - c = c - ch2oh% Rfi in which formulas Rg and Rg are separately selected from a group consisting of hydrogen and (CH2) pOH, where p is 1-4, provided that at least one of the groups Rg and Rg is hydrogen, or together Rg and Rg together form a carbon-carbon supplemental bond, such that an alkyne is uprooted, and its physiologically suitable salts ooh geometric isomers. 4. A process according to claim 1 or 3, characterized in that R 1 and R 5 are hydroxymethyl of formulas I and III. Process according to claim 1 or 3, characterized in that the compound 9- [4-hydroxy-3- (hydroxymethyl) -2-butenyl] -guanine is prepared. Process according to claim 1 or 3, characterized in that the compound 9- (4-hydroxy-2-butenyl) guanine is prepared. Process according to claim 6, characterized in that the compound cis-9- (4-hydroxy-2-butenyl) guanine is prepared. Process according to claim 1 or 3, characterized in that the compound 9- (4-hydroxy-2-butynyl) guanine is prepared. 9. A process according to claim 1 or 2, characterized in that the compound 9- [4-hydroxy-2- (hydroxymethyl) butyl] guanine is prepared. Process according to claims 1, 2 or 9, characterized in that the compound produced is in the form of an optical isomer. Process according to any one of claims 1, 3-6, characterized in that the compound produced is in the form of a geometric isomer. Process according to any one of claims 1-11, characterized in that the compound produced is in the form of a physiologically suitable site.